视黄醇对LPS诱发的原代培养大鼠乳腺上皮细胞炎症反应的调节

本研究旨在优化大鼠乳腺上皮细胞培养体系,建立LPS诱发的炎症反应模型,研究视黄醇对其炎症反应的调节机制.采用胶原酶与透明质酸酶联合消化分离大鼠乳腺上皮细胞.待细胞铺满整个培养瓶的90％时,(1)用不同浓度的LPS处理乳腺上皮细胞,24 h收集细胞及培养上清液；(2)分为试验组(添加1 μmol· L-1视黄醇)和对照组,处理24 h后更换培养液,用10μg·mL-1的LPS处理细胞,分别于不同时间点收集细胞.结果表明:LPS 处理大鼠乳腺上皮细胞后引起各个时间点炎性因子mRNA表达极显著升高,视黄醇能显著下调上述炎性细胞因子的表达.视黄醇预处理能减轻LPS诱发的大鼠乳腺上皮细胞的炎症损伤.

Modulation of Retinoic Acid in LPS-induced Inflammation in Primary Culture of Rat Mammary Epithelial Cells

Lipopolysaccharide(LPS) induced primary culture of rat mammary epithelial cells(rMEC) inflammatory model was established and the protective mechanism of retinoic acid was studied.rMEC s were separated through digestion by collagenase and hyaluronidase.When cells were grown to 90% confluence in 6-well plates,(1) cells were treated with different concentrations of LPS;(2) cells were treated with 1 μmol·L-1 retinoic acid before stimulation with 10 μg· mL-1 LPS.Cells and supernatants were collected.In this study,inflammatory models of primary culture of rMEC were successfully established.Stimulation of rMEC with LPS elicited a marked increase in mRNA expression for inflammatory mediators.Treatment of retinoic acid significantly decreased the mRNA expression of those mediators.Retinoic acid could reduce LPS-induced inflammatory reaction.These results further indicate that retinoic acid may protect mastitis from pro-inflammatory cytokine-mediated damage of mammary epithelial cells.